家蚕抗菌肽Attacin基因植物表达载体的构建

抗菌肽是昆虫抵御外界微生物侵染的主要物质。利用双酶切将pBI121载体上的植物启动子CaMV35S克隆到植物表达载体pCAMBIA2300上,构建重组表达载体pCAMBIA2300-CaMV35S。通过PCR扩增家蚕抗菌肽attacin基因编码区全长序列,并将其成功克隆至T载体上(Gen Bank登录号:GU244351),然后利用双酶切将attacin基因亚克隆到pCAMBIA2300-CaMV35S上,通过PCR鉴定,成功构建了attacin基因的植物表达载体pCAMBIA2300-Ca MV35S-attacin。为研究attacin基因在植物抗病性方面的应用奠定基础。

Construction of Plant Expression Vector with Attacin in Silkworm, Bombyx mori.

Antimicrobial peptides are the most important substance for insect in protecting themselves from infection of microorganism. In the paper, the plant-specific promoter CaMV35S from pBI121 was constructed into pCAMBIA2300, the recombinant expression vector named pCAMBIA2300-CaMV35S, attacin full-length coding region (GenBank ccession number: GU244351) as amplified by PCR from Bombyx mori and then cloned into the pMD19-T simple vector. The positive clone cutted by pairs of restriction enzymes was successfully linked with recombinant expression vector pCAMBIA2300- CaMV35S. The recombinat plasmid of attacin was identified by PCR digestion. That lay the foundation for applications of the antibacterial peptides attacin gene in disease resistance of plant.